beta Actin Rabbit Polyclonal Antibody

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Catalog# R1207-1

beta Actin Rabbit Polyclonal Antibody

Application

WB
IF-Cell
IHC-P
FC

Reactivity

Human
Mouse
Rat
Zebrafish
bamboo

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

beta Actin Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within N-terminal residues of β-Actin.

Species Reactivity

Human, Mouse, Rat, Zebrafish, bamboo

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 42 kDa

Positive Control

HeLa cell lysate, A549 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, mouse lung tissue lysate, rat lung tissue lysate, HeLa, NIH/3T3, PC-12, human kidney tissue, human lung tissue, mouse kidney tissue, mouse lung tissue, rat kidney tissue, rat lung tissue, hybrid fish (crucian-carp) brain tissue, hybrid fish (crucian-carp) kidney tissue.

Conjugation

unconjugated

RRID

AB_3073201

Product Features

Form

Liquid

Concentration

1ug/ul

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

WB
1:20,000

IF-Cell
1:100

IHC-P
1:2,000

FC
1:1,000

Applications in Publications

WB	See 83 publications below
IF	See 2 publications below
IHC	See 1 publications below

Species in Publications

Mouse	See 33 publications below
Human	See 30 publications below
Zebrafish	See 5 publications below

Target

Function

Actins are highly conserved proteins involved in cell motility, structure and integrity. Actin has been found to be expressed in at least six isomeric forms. It is expressed in heart and skeletal striated muscle tissue, and in certain smooth muscle tissues, regulating contractile potentials for these cells. It is also expressed in the cytoplasm of non-muscle cells, functioning to control cell structure and motility. Beta actin is usually used as a loading control, for among others, the integrity of cells, protein degradation, in Western Blotting.

Background References

1. Ponte P., Ng S.Y., Engel J., Gunning P., Kedes L."Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human beta-actin cDNA." Nucleic Acids Res. 12:1687-1696(1984) Ohmori H., Toyama S., Toyama S."Direct proof that the primary site of action of cytochalasin on cell motility processes is actin."J. Cell Biol. 116:933-941(1992)

Sequence Similarity

Belongs to the actin family.

Post-translational Modification

ISGylated.; Oxidation of Met-44 and Met-47 by MICALs (MICAL1, MICAL2 or MICAL3) to form methionine sulfoxide promotes actin filament depolymerization. MICAL1 and MICAL2 produce the (R)-S-oxide form. The (R)-S-oxide form is reverted by MSRB1 and MSRB2, which promote actin repolymerization.; Monomethylation at Lys-84 (K84me1) regulates actin-myosin interaction and actomyosin-dependent processes. Demethylation by ALKBH4 is required for maintaining actomyosin dynamics supporting normal cleavage furrow ingression during cytokinesis and cell migration.; Methylated at His-73 by SETD3. Methylation at His-73 is required for smooth muscle contraction of the laboring uterus during delivery (By similarity).; [Actin, cytoplasmic 1, N-terminally processed]: N-terminal acetylation by NAA80 affects actin filament depolymerization and elongation, including elongation driven by formins. In contrast, filament nucleation by the Arp2/3 complex is not affected.; (Microbial infection) Monomeric actin is cross-linked by V.cholerae toxins RtxA and VgrG1 in case of infection: bacterial toxins mediate the cross-link between Lys-50 of one monomer and Glu-270 of another actin monomer, resulting in formation of highly toxic actin oligomers that cause cell rounding. The toxin can be highly efficient at very low concentrations by acting on formin homology family proteins: toxic actin oligomers bind with high affinity to formins and adversely affect both nucleation and elongation abilities of formins, causing their potent inhibition in both profilin-dependent and independent manners.

Subcellular Location

Cytoskeleton

UNIPROT

SwissProt: P60709 Human

SwissProt: P60710 Mouse

SwissProt: P60711 Rat

SwissProt: Q7ZVI7 Zebrafish

Synonyms

A26C1A antibody

A26C1B antibody

ACTB antibody

ACTB_HUMAN antibody

Actin beta antibody

Actin cytoplasmic 1 antibody

Actin, cytoplasmic 1, N-terminally processed antibody

Actx antibody

b actin antibody

Beta cytoskeletal actin antibody

Expand

Images

Western blot analysis of beta Actin on different lysates with Rabbit anti-beta Actin antibody (R1207-1) at 1/20,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: Jurkat cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: Mouse brain tissue lysate
Lane 7: Rat brain tissue lysate
Lane 8: Mouse lung tissue lysate
Lane 9: Rat lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 30 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (R1207-1) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HeLa cells labeling beta Actin with Rabbit anti-beta Actin antibody (R1207-1) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta Actin antibody (R1207-1) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of NIH/3T3 cells labeling beta Actin with Rabbit anti-beta Actin antibody (R1207-1) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta Actin antibody (R1207-1) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of PC-12 cells labeling beta Actin with Rabbit anti-beta Actin antibody (R1207-1) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta Actin antibody (R1207-1) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-beta Actin antibody (R1207-1) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1207-1) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-beta Actin antibody (R1207-1) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1207-1) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-beta Actin antibody (R1207-1) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1207-1) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-beta Actin antibody (R1207-1) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1207-1) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-beta Actin antibody (R1207-1) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1207-1) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-beta Actin antibody (R1207-1) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (R1207-1) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western blot analysis of β-actin on different lysates using anti-β-actin antibody at 1/1,000 dilution.
Positive control:
Lane 1: hybrid fish (crucian-carp) brain tissue
Lane 2: hybrid fish (crucian-carp) kidney tissue